NDLI: Optimization of a FACS based-immunoassay using E. coli autodisplaying Z-domains

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BioChip Journal : Volume 7, Issue 1, March 2013

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Optimization of a FACS based-immunoassay using E. coli autodisplaying Z-domains

Content Provider	Springer Nature Link
Author	Park, Min Bong, Ji Hong Yoo, Gu Jose, Joachim Kang, Min Jung Pyun, Jae Chul
Copyright Year	2013
Abstract	We developed a fluorescence-activated cell sorter (FACS)-based immunoassay using Escherichia coli with autodisplayed Z-domains as a solid support for detection antibodies as well as a method to optimize the signal-to-noise ratio of fluorescence signals from E. coli. To improve the sensitivity of FACS measurements by eliminating artifactual signals from unrelated small particles in the sample mixture, E. coli with autodisplayed Z-domains was modified to express enhanced green fluorescence protein (eGFP) in the cytosol. By selectively gating the measurement channel for eGFP in the cytosol, we were able to discriminate between green fluorescent E. coli and artifact particles. We confirmed the efficacy of this optimization method for FACS-based immunoassay using several model reactions. Finally, as proof of concept, we used the FACS-based immunoassay and our optimization method for the medical diagnosis of human hepatitis B virus surface antigen.
Starting Page	173
Ending Page	179
Page Count	7
File Format	PDF
ISSN	19760280
Journal	BioChip Journal
Volume Number	7
Issue Number	2
e-ISSN	20927843
Language	English
Publisher	Springer-Verlag
Publisher Date	2013-06-20
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Immunoassay FACS Z-domain Medical diagnosis Hepatitis B virus surface antigen Biotechnology Biomedical Engineering
Content Type	Text
Resource Type	Article
Subject	Bioengineering Biomedical Engineering Electrical and Electronic Engineering Biotechnology

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3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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